CN114223729A - Application of bifidobacterium longum subsp. infantis and breast milk oligosaccharide composition in formula milk powder - Google Patents

Application of bifidobacterium longum subsp. infantis and breast milk oligosaccharide composition in formula milk powder Download PDF

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CN114223729A
CN114223729A CN202111447747.8A CN202111447747A CN114223729A CN 114223729 A CN114223729 A CN 114223729A CN 202111447747 A CN202111447747 A CN 202111447747A CN 114223729 A CN114223729 A CN 114223729A
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formula
bifidobacterium longum
infantis
breast milk
staphylococcus aureus
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CN114223729B (en
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郝威
刘彪
李威
孔小宇
肖竞舟
吉塞拉·阿德里安娜·怀斯
卡洛琳·安妮卡·凡·鲁-鲍曼
盖瑞特·施密特
王雯丹
司徒文佑
刘伟贤
洪维鍊
王燕霞
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Inner Mongolia Yili Industrial Group Co Ltd
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Inner Mongolia Yili Industrial Group Co Ltd
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C21/00—Whey; Whey preparations
    • A23C21/06—Mixtures of whey with milk products or milk components
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C21/00—Whey; Whey preparations
    • A23C21/02—Whey; Whey preparations containing, or treated with, microorganisms or enzymes
    • A23C21/026—Whey; Whey preparations containing, or treated with, microorganisms or enzymes containing, or treated only with, lactic acid producing bacteria, bifidobacteria or propionic acid bacteria
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C21/00—Whey; Whey preparations
    • A23C21/04—Whey; Whey preparations containing non-milk components as source of fats or proteins
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C21/00—Whey; Whey preparations
    • A23C21/08—Whey; Whey preparations containing other organic additives, e.g. vegetable or animal products
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C21/00—Whey; Whey preparations
    • A23C21/10—Whey; Whey preparations containing inorganic additives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/51—Bifidobacterium
    • A23V2400/531—Lactis
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/51—Bifidobacterium
    • A23V2400/533—Longum

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  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
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  • Inorganic Chemistry (AREA)
  • Microbiology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Dairy Products (AREA)

Abstract

The invention provides application of a bifidobacterium longum subsp. infantis and breast milk oligosaccharide composition in formula milk. Specifically, the invention provides an application of a composition of bifidobacterium longum subsp. infantis and breast milk oligosaccharide in preparing formula milk powder which can effectively improve the resistance of organisms to staphylococcus aureus infection and can improve the innate immunity and/or anti-aging effect of the organisms, wherein each 100g of the formula milk powder contains 1 x 10 of the bifidobacterium longum subsp. infantis3CFU～1×1012CFU, 0.1g to 10g of breast milk oligosaccharide.

Description

Application of bifidobacterium longum subsp. infantis and breast milk oligosaccharide composition in formula milk powder
Technical Field
The invention relates to an application of a bifidobacterium longum subsp. infantis and breast milk oligosaccharide composition in preparing formula milk powder.
Background
Over the last millennium, the medical literature has documented that non-breastfed infants have a higher rate of illness and mortality than breastfed infants. The breast milk not only provides the required nutrition for the infant, but also provides the guarantee for the intestinal development and the immunity improvement of the infant by the active ingredients in the breast milk. Breast-fed infants have a higher relative abundance of beneficial bacteria, particularly bifidobacteria and lactic bacteria, in the gut flora compared to formula-fed infants.
Transmission of breast milk by floraIn addition, the active ingredients such as breast milk oligosaccharide and cytokine in breast milk are added to establish healthy intestinal flora for the newborn. The infant intakes 10 via breast milk every day7-108Individual bacteria, including lactic acid bacteria and bifidobacteria. The bacteria are directly transmitted to the infant through breast milk, and part of the bacteria can be planted in the intestinal tract of the infant, so that the establishment of the intestinal flora in the early life is promoted. The establishment of the infant's intestinal flora has short-term, even lifelong effects on the development of its intestinal tract, as well as on the health and immune system.
Human Milk Oligosaccharides (HMOs) are the third largest group of emulsion compounds present in Human Milk. It plays an important role in early infant development. To date, many different structures have been identified 130, but the functional impact is not known. Most oligosaccharides in milk cannot be digested and therefore HMOs can act as prebiotics. Furthermore, HMO in the maturation of the immune system and its role as an immunomodulator shows its importance for the healthy development of infants. Currently, HMOs have been added to infant formulas.
At present, in the fields of infant formula powder, supplementary food and nutritional supplements, a solution for relieving infant intestinal discomfort and improving the self-resistance to pathogenic bacteria such as staphylococcus aureus is needed. Meanwhile, in the field of children, teenagers and adults over 3 years old, solutions for alleviating intestinal discomfort and improving the self-defense against pathogenic bacteria such as staphylococcus aureus infection are also needed.
Caenorhabditis elegans (C.elegans) has a good application prospect in preclinical research and evaluation as a model organism. It has short life cycle (21 days), strong reproducibility and regeneration, simple operation, transparency and easy culture. Its genome has been completely sequenced and one quarter of the genes are homologous to the human genome. The nematode organism with gene mutation produced by editing nematode gene can be used as experimental means for gene analysis. Nematodes are not currently considered an animal in european legislation. It is widely used as an in vitro assay such as transcriptomics, proteomics, metabolomics, and the like. As a model organism, it is also often used as the first step in the evaluation of materials, often using nematodes as a high throughput means to screen test materials for certain properties prior to design of functional materials, in vitro enzymatic or cellular assays, mouse models and clinical trials.
Staphylococcus aureus (s. aureus) is a gram-positive bacterium belonging to the genus Staphylococcus, a common food-borne pathogenic microorganism. It is widely present in natural environment, and under proper conditions, it can produce enterotoxin and cause food poisoning, which causes food-borne microbial food poisoning events caused by staphylococcus aureus. After a human body is infected by staphylococcus aureus, common food poisoning symptoms such as nausea, vomiting, dizziness and the like can appear, and symptoms such as enteritis, pneumonia, skin infection, wound infection fester, meningitis and the like can appear.
Disclosure of Invention
The inventor of the present invention finds in research that the combination of some bifidobacterium longum subspecies infants and breast milk oligosaccharides can improve the capability of organisms to resist staphylococcus aureus infection, thereby providing the following invention:
in one aspect, the present invention provides the use of a composition of bifidobacterium longum subsp. infantis and breast milk oligosaccharides in the preparation of a milk formula having the effects of effectively improving the resistance of an organism against staphylococcus aureus infection and improving the innate immunity and/or anti-aging of the organism, wherein 100g of the milk formula contains 1 × 10 bifidobacterium longum subsp. infantis3CFU～1×1012CFU/100g, breast milk oligosaccharide 0.1-10 g.
On the other hand, the invention also provides the formula milk powder which can effectively improve the resistance of organisms to staphylococcus aureus infection and improve the innate immunity and/or anti-aging effect of the organisms, and the formula milk powder is prepared by adding bifidobacterium longum subspecies infants and breast milk oligosaccharide into the formula milk powder.
According to a particular embodiment of the invention, said Bifidobacterium longum subsp. infantis strain in said formula of the invention comprises Bifidobacterium longum subsp. infantis strain YLGB-1496 with a accession number CGMCC No. 21109. Bifidobacterium longum subspecies of infantis with the preservation number of CGMCC No.21109 has been preserved in China general microbiological culture Collection center (CGMCC) at 12 months and 05 days of 2020, and the preservation unit address is as follows: west road No. 1, north west of the morning area, beijing, institute of microbiology, china academy of sciences, date of deposit: year 2020, 12 months and 05 days; the preservation number is: CGMCC No. 21109; and (3) classification and naming: bifidobacterium longum subsp. The strain is also called YLGB-1496 or GB-1496 in the invention.
According to a specific embodiment of the invention, in the formula milk powder, the total protein content is 10-20 g/100g, and the proportion of whey protein in the total protein is 40-70%; the fat content is 15 g-30 g/100 g; the carbohydrate content is 50 g-70 g/100 g.
According to a specific embodiment of the invention, the formula milk powder further comprises linoleic acid and alpha-linolenic acid, wherein the content of the linoleic acid is 2700-4500 mg/100g, and the content of the alpha-linolenic acid is 270-450 mg/100 g.
According to a specific embodiment of the present invention, the formula of the present invention may be an infant formula (infant formula) or a non-infant formula (e.g. children formula, adult formula).
According to a specific embodiment of the present invention, the raw materials for providing total protein in the formula of the present invention comprise: raw milk (cow/sheep), whole milk powder (cow/sheep), skimmed milk powder (cow/sheep), whey protein powder (cow/sheep), desalted whey powder (cow/sheep), and one or more of beta-casein. Specifically, based on 1000 parts by weight of the formula milk powder, the formula milk powder comprises the following raw materials: 0-3500 parts of raw milk (cow/sheep), 0-260 parts of whole milk powder (cow/sheep), 0-120 parts of whey protein powder (cow/sheep), 0-400 parts of skimmed milk powder (cow/sheep), and/or 0-600 parts of desalted whey powder (cow/sheep), and 0-40 parts of beta-casein.
According to a specific embodiment of the present invention, in the formula of the present invention, the raw material for providing fat comprises one or more of the following raw materials in parts by weight: 0-50 parts of corn oil, 0-65 parts of soybean oil, 0-100 parts of sunflower seed oil, 0-240 parts of 1, 3-dioleoyl-2-palmitic triglyceride, 0-30 parts of coconut oil, 0-85 parts of low erucic acid rapeseed oil and 0-20 parts of linseed oil.
According to a specific embodiment of the present invention, the raw materials for providing carbohydrates in the formula of the present invention comprise: 0-500 parts of lactose and/or the protein raw material containing lactose.
According to a particular embodiment of the invention, the formula of the invention may comprise, in addition to the above-mentioned components, the usual components of formula, such as one or more of nutrients, dietary fibres, inositol, taurine, L-carnitine, docosahexaenoic acid, arachidonic acid, lutein, nucleotides, lactoferrin. Preferably comprises a compound nutrient bag. In some preferred embodiments, the formula comprises 0-20 parts of nutrients including vitamins and minerals, or further optionally one or more of dietary fiber (fructooligosaccharides, galactooligosaccharides, polyfructose, etc.), inositol, taurine, L-carnitine, docosahexaenoic acid, arachidonic acid, lutein, nucleotides, lactoferrin. The compound nutrient mainly comprises: compounding vitamins: vitamin A, vitamin D, vitamin E, and vitamin K1Vitamin B1Vitamin B2Vitamin B6Vitamin B12Nicotinic acid, folic acid, pantothenic acid, vitamin C, choline and biotin; compounding minerals: sodium, potassium, calcium, phosphorus, copper, iron, zinc, manganese, iodine, selenium, magnesium, potassium and derivatives thereof. The source, content and usage amount of the vitamins and minerals should meet the national standards and relevant regulations of infant formula food.
According to a particular embodiment of the invention, the present invention is applied in a composition of bifidobacterium longum subspecies infants and breast milk oligosaccharides for the preparation of a formula, said breast milk oligosaccharides comprising one or more of 2 '-fucosyllactose, 3' -fucosyllactose, lacto-N-tetraose, 3 '-sialyllactose, 6' -sialyllactose.
2 ' -fucosyllactose (2 ' -fucosyllactose, 2 ' -FL) is a trisaccharide structure formed by fucose and lactose, and is a representative substance of fucosyl oligosaccharides. The commercially available material is usually prepared by microbial fermentation and has the same structure as 2' -fucosyllactose found in human milk.
3-fucosyllactose (3-fucosyllactose, 3-FL) is a trisaccharide structure formed by fucose and lactose, is an isomer of 2' -fucosyllactose, and is a representative substance of fucosyl oligosaccharides. This material is generally commercially available as prepared by microbial fermentation and has the same structure as 3-fucosyllactose found in human milk.
lacto-N-tetraose (LNT), a hexasaccharide structure formed by lactose and tetraose, is a representative substance of oligosaccharides having a core sugar chain as a basic structure and not containing a fucosyl group or a sialyl group, which is generally commercially available through a microbial fermentation process and has the same structure as lacto-N-tetraose found in human milk.
3 ' -sialyllactose (3 ' -SL) is a trisaccharide structure formed by sialic acid and lactose, and is a representative substance of sialic acid-based oligosaccharides, as an isomer with 6 ' -sialyllactose. This material is generally commercially available as prepared by microbial fermentation and has the same structure as 3' -sialyllactose found in human milk.
6 '-sialyllactose (6' -sialyllactose, 6 '-SL) is a trisaccharide structure formed by sialic acid and lactose, and is a representative substance of sialic acid-based oligosaccharides as an isomer with 3' -sialyllactose. This material is generally commercially available as prepared by microbial fermentation and has the same structure as 6' -sialyllactose found in human milk.
According to a particular embodiment of the invention, the present invention is applied to the composition of bifidobacterium longum subsp. The content of 2' -fucosyllactose in the breast milk oligosaccharide may be 80% or more, for example 85% or more, 90% or more, 95% or more, or 100%.
According to a particular embodiment of the invention, the present invention is applied in a composition of bifidobacterium longum subspecies infantis and breast milk oligosaccharides for the preparation of a formula, said breast milk oligosaccharides being a combination consisting of at least two, at least three or at least four of 2 '-fucosyllactose (2' -FL), 3-fucosyllactose (3-FL), lacto-N-tetraose (LNT), 3 '-sialyllactose (3' -SL) and 6 '-sialyllactose (6' -SL).
According to a particular embodiment of the invention, the present invention is applied in a composition of bifidobacterium longum subspecies infantis and breast milk oligosaccharides for the preparation of a formula wherein the ratio of bifidobacterium longum subspecies infantis to breast milk oligosaccharides is 1 x 103CFU～1×1012CFU: 0.1g to 10g, preferably 1X 106CFU～1×1010CFU: 1g to 10 g. In some more specific embodiments of the invention, the ratio of bifidobacterium longum subspecies infantis to breast milk oligosaccharides is 1 x 108CFU: 0.08g to 0.3 g. The ratio of bifidobacterium longum subsp. infantis to breast milk oligosaccharides is based on the amount of both in the same composition.
According to a specific embodiment of the present invention, the use of the composition of the present invention for enhancing the resistance of an organism against staphylococcus aureus infection comprises: improving the ability of an individual to prevent staphylococcus aureus infection, reducing the ability of an individual to be infected with staphylococcus aureus, and/or relieving various symptoms caused by staphylococcus aureus infection, such as food poisoning (including nausea, vomiting, dizziness and the like), enteritis, pneumonia, skin infection, wound ulceration or meningitis and the like.
According to a particular embodiment of the invention, for use of the composition of the invention, the organism comprises an animal or a human.
The formula milk powder provided by the invention has the effects of effectively improving the resistance of organisms to staphylococcus aureus infection and improving the innate immunity and/or anti-aging effect due to the composition comprising the bifidobacterium longum subspecies infantis and breast milk oligosaccharide.
In another aspect, the present invention also provides a method for preparing the milk formula, wherein the method comprises the step of mixing the bifidobacterium longum subsp. infantis and breast milk oligosaccharide composition with other raw materials in the formula by adopting a wet or dry production process to prepare the breast emulsified milk formula. The preparation process mainly comprises the following steps: preparing materials, homogenizing, concentrating, sterilizing, spray drying, and dry mixing to obtain the final product. In the present invention, bifidobacterium longum subspecies infantis may be mixed together at the time of compounding (in inactivated form in the infant formula), preferably added during post-mixing (in viable form in the infant formula). In the invention, the breast milk oligosaccharide can be mixed together during the compounding process, and can also be added in the post-mixing process.
In conclusion, the composition comprising bifidobacterium longum subsp. infantis and breast milk oligosaccharide is found to have the effects of resisting aging, improving the innate immunity of organisms and effectively improving the resistance of the organisms to staphylococcus aureus infection, can resist infection and improve the immunity when being applied to formula milk powder, and has wide application prospect.
Drawings
FIG. 1 shows the effect of a 15mg/mL combination of the breast milk oligosaccharide 2' -FL and the probiotic YLGB-1496 on the survival rate of C.elegans when infected with S.aureus.
Microbial deposits for patent procedures:
bifidobacterium infantis YLGB-1496 (or GB-1496) strain:
the preservation date is as follows: year 2020, 12 months and 05 days;
the preservation unit: china general microbiological culture Collection center (CGMCC);
the address of the depository: western road No. 1, north west city of township, beijing, institute of microbiology, china academy of sciences;
the preservation number is: CGMCC No. 21109;
and (3) classification and naming: bifidobacterium longum subsp.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments that can be derived by one of ordinary skill in the art from the embodiments given herein are intended to be within the scope of the present invention.
Unless specifically defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the relevant art.
The operating conditions not specified in detail in the examples were carried out according to the usual procedures in the art.
The probiotic Bifidobacterium longum subsp. infantis YLGB-1496 (the source is the breast milk of Chinese healthy mother, which is preserved in China general microbiological culture Collection center (CGMCC) in 12 months and 05 days in 2020, the preservation unit address is No. 3 of West Lu No. 1 of Beijing south China area facing the sun, the preservation date is No. 05 days in 2020, the preservation number is CGMCC No.21109, and the classification name is Bifidobacterium longum subsp. infantis) used in the invention.
The HMO raw materials used in the invention are all from Jennewein.
Example 1 infant formula containing probiotic and prebiotic composition (1000 kg prepared):
1000 kg of raw milk, 320 kg of lactose, 25 kg of whey protein powder WPC 80%, 90175 kg of desalted whey powder D, 40 kg of corn oil, 50 kg of soybean oil, 140 kg of OPO structural fat, 27 kg of alpha-whey protein powder, 9 kg of beta-casein powder, 1 kg of anhydrous cream, 17 kg of fructo-oligosaccharide powder, 40 kg of galacto-oligosaccharide syrup, 2 kg of breast milk oligosaccharide (2' -FL), 17 kg of compound nutrient, 12 kg of DHA, 22 kg of ARA, YLGB-14960.1 kg of bifidobacterium longum subspecies and 0.1 kg of bifidobacterium lactis.
The compound nutrient comprises about 2.5 kg of compound vitamin nutrient package, about 0.75 kg of choline chloride nutrient package, about 6 kg of calcium powder nutrient package, about 1 kg of mineral nutrient package, about 1.5 kg of magnesium chloride nutrient package and about 2 kg of potassium chloride nutrient package, and the base material of each nutrient package is lactose.
The preparation process of the infant formula containing the probiotic and prebiotic composition comprises the following steps:
1) milk rough filtration: after coarse filtration and degassing in a balance cylinder, the milk is preheated by a plate heat exchanger, and impurities are separated by a separator.
2) Homogenizing and sterilizing milk: one part of the raw milk without impurities enters a homogenizer for homogenization, the other part of the raw milk is inhomogeneous, and the homogenized raw milk are mixed and enter a sterilization system for sterilization.
3) Adding powder: various powder raw materials are metered according to the formula, uniformly added into a powder preparation tank through an air conveying system, and sucked into a vacuum mixing tank through a vacuum system;
4) dissolving and oil blending: putting the grease specified in the formula into an oil dissolving chamber according to the formula requirement, keeping the temperature of the oil dissolving chamber at 50-90 ℃, pouring the oil into a mixed oil storage tank after the oil is dissolved, and pouring the mixed oil into a material mixing tank through an oil pump according to the formula requirement;
5) dissolving and adding nutrients: respectively dissolving nutrient bags such as calcium powder, vitamins, minerals, etc. with purified water, and sequentially adding into a mixing tank to obtain mixed feed liquid.
6) Dissolving and adding breast milk oligosaccharide: and (3) dissolving the breast milk oligosaccharide raw material by using part of the mixed material liquid in the step (5), and adding the dissolved breast milk oligosaccharide raw material into a mixing tank to obtain the mixed material liquid containing 2' FL.
7) And (3) filtering: filtering the mixed feed liquid containing the breast milk oligosaccharide by a filter screen to remove physical impurities possibly brought in the raw materials.
8) Homogenizing: homogenizing the mixed material liquid by a homogenizer, mechanically processing the fat balls, and dispersing the fat balls into uniform fat balls.
9) Cooling and storing: and (3) feeding the homogenized material liquid into a plate heat exchanger for cooling: cooling to below 20 ℃, temporarily storing in a pre-storage cylinder, entering the next procedure within 6 hours, and starting the stirrer according to the set requirement.
10) Concentration and sterilization: double-effect concentration is adopted during production, the sterilization temperature is more than or equal to 83 ℃, and the sterilization time is 25 seconds. The discharge concentration was 50% dry matter.
11) Storing concentrated milk, preheating, filtering and spray drying: the concentrated milk is temporarily stored in a concentrated milk balancing tank. Preheating to 60 deg.C with a scraper preheater, filtering the preheated material with a filter with 1mm pore diameter, pumping into a drying tower with a high pressure pump, spray drying, and agglomerating fine powder at the tower top or fluidized bed as required. Air inlet temperature: 180 ℃, the exhaust temperature is 86 ℃, the pressure of the high-pressure pump is 200bar, and the negative pressure of the tower is about-4 mba.
12) Drying and cooling the fluidized bed: and (3) drying the powder from the drying tower for the second time by the fluidized bed (the first stage), and cooling to 30 ℃ by the fluidized bed (the second stage) to obtain the main milk powder material.
13) Subpackaging: according to the formula requirements, DHA, ARA or bifidobacterium longum subspecies YLGB-1496 or bifidobacterium lactis are weighed, sealed and packaged.
14) Dry mixing: the weighed DHA, ARA or Bifidobacterium longum subspecies YLGB-1496 and the milk powder main material are evenly mixed in a dry mixer.
15) Powder sieving: the granularity of the milk powder is uniform through the vibrating screen, and the powder residue is discarded.
16) Powder discharging: and (4) receiving the powder by using a sterilized powder collecting box, and conveying the powder to a powder feeding room from a powder discharging room.
17) Powdering: pouring the milk powder into a powder storage tank on a large and small packaging machine according to the packaging requirements.
18) Packaging: 400 g of the mixture is packaged by an automatic packaging machine in a nitrogen-filled mode. The oxygen content is lower than 1% when charging nitrogen. The oxygen content of the 900 g iron can in the automatic nitrogen-filled package is lower than 5 percent.
19) Boxing: and (4) filling the packaged small bags into a paper box, adding a powder spoon, and sealing by using a box sealing machine.
20) And (4) inspecting a finished product: and sampling and inspecting the packaged product according to an inspection plan.
21) And (4) warehousing and storing: and warehousing and storing the qualified product at normal temperature with the humidity less than or equal to 65 percent.
In the product, the content of breast milk oligosaccharide 2' FL is about 0.2g/100 g; bifidobacterium longum subspecies YLGB-1496 content is 1 × 106CFU/g。
Example 2 infant formula containing probiotic and prebiotic composition (1000 kg prepared):
1000 kg of raw milk, 250 kg of skim milk powder, 150 kg of lactose, 50 kg of whey protein powder WPC 34%, 50 kg of desalted whey powder D90225 kg, 106 kg of OPO structural fat, 37 kg of soybean oil, 30 kg of corn oil, 10 kg of alpha-whey protein powder, 10 kg of beta-casein powder, 5kg of fructo-oligosaccharide powder, 15 kg of galacto-oligosaccharide syrup, 4 kg of soybean lecithin, 11 kg of compound nutrient, 12 kg of DHA, 14 kg of ARA, 8kg of breast milk oligosaccharide (2' FL), LGYB-14960.2 kg of bifidobacterium longum subspecies and 0.65 kg of nucleotide.
The compound nutrient comprises about 1.5 kg of compound vitamin nutrient package, about 0.75 kg of choline chloride nutrient package, about 5kg of calcium powder nutrient package, about 1 kg of mineral nutrient package, about 0.75 kg of magnesium chloride nutrient package and about 2 kg of potassium chloride nutrient package, and the base material of each nutrient package is lactose. The product preparation process was as in example 1.
In the product, the content of breast milk oligosaccharide 2' FL is about 0.8g/100 g; bifidobacterium longum subspecies YLGB-1496 content is 2 × 106CFU/g。
Example 3 infant formula containing probiotic and prebiotic composition (1000 kg prepared):
1000 kg of raw milk, 250 kg of skim milk powder, 140 kg of lactose, 50 kg of whey protein powder WPC 34%, 50 kg of desalted whey powder D90225 kg, 106 kg of OPO structural fat, 37 kg of soybean oil, 30 kg of corn oil, 10 kg of alpha-whey protein powder, 10 kg of beta-casein powder, 5kg of fructo-oligosaccharide powder, 15 kg of galacto-oligosaccharide syrup, 30 kg of breast milk oligosaccharide (2' FL), 11 kg of compound nutrient, 12 kg of DHA, 14 kg of ARA, YLGB-14960.4 kg of bifidobacterium longum subspecies and 0.65 kg of nucleotide.
The compound nutrient comprises about 1.5 kg of compound vitamin nutrient package, about 0.75 kg of choline chloride nutrient package, about 5kg of calcium powder nutrient package, about 1 kg of mineral nutrient package, about 0.75 kg of magnesium chloride nutrient package and about 2 kg of potassium chloride nutrient package, and the base material of each nutrient package is lactose. The product preparation process was as in example 1.
In the product, the content of breast milk oligosaccharide 2' FL is about 3g/100 g; bifidobacterium longum subspecies YLGB-1496 content is 4 × 106CFU/g。
Example 4 infant formula containing probiotic and prebiotic composition (1000 kg prepared):
1000 kg of raw milk, 250 kg of skim milk powder, 165 kg of lactose, 50 kg of whey protein powder WPC 34%, 50 kg of desalted whey powder D90225 kg, 106 kg of OPO structural fat, 37 kg of soybean oil, 30 kg of corn oil, 10 kg of alpha-whey protein powder, 10 kg of beta-casein powder, 5kg of fructo-oligosaccharide powder, 15 kg of galacto-oligosaccharide, 5kg of breast milk oligosaccharide (2' FL 2.65kg and LNT 0.8kg), 11 kg of compound nutrient, 12 kg, 14 kg of ARA, 11 kg of Bifidobacterium longum subspecies YLGB-14960.4 kg and 0.65 kg of nucleotide.
The compound nutrient comprises about 1.5 kg of compound vitamin nutrient package, about 0.75 kg of choline chloride nutrient package, about 5kg of calcium powder nutrient package, about 1 kg of mineral nutrient package, about 0.75 kg of magnesium chloride nutrient package and about 2 kg of potassium chloride nutrient package, and the base material of each nutrient package is lactose. The product preparation process was as in example 1.
In the product, the content of the 2' FL in the breast milk oligosaccharide composition is about 0.265g/100g, and the content of the LNT is about 0.8g/100 g; bifidobacterium longum subspecies YLGB-1496 content is 4 × 106CFU/g。
Efficacy experiment of Bifidobacterium longum subspecies infants and breast milk oligosaccharide composition
Probiotic bifidobacterium longum subsp. infantis YLGB-1496 the probiotic bacteria tested were cultured in MRS medium supplemented with 1% cysteine and incubated overnight at 37 ℃ in an anaerobic environment. The thallus is harvested and washed with normal saline solution, and inoculated into plates containing nematode growth medium with probiotic concentration of 1 × 10 per plate8CFU or 1X 107CFU。
HMOs use distilled water to formulate breast milk oligosaccharide solutions.
Nematode infection model
Nematodes of consistent age were obtained and cultured in dishes containing nematode agar medium (NGM medium, containing E.coli OP50 as food) supplemented with different doses of HMO (1)5mg/mL) and probiotic (1X 10)8CFU、1×107CFU) composition was co-cultured (total volume of liquid added per nematode petri dish was 10 mL. For the HMO test group at a concentration of 15mg/mL, the final amount of HMO per dish was 150 mg. Therefore, the ratio of probiotic to HMO added in each plate was: 1X 108CFU: 150mg, or 1X 107CFU: 150 mg). After the nematodes became adult, they were transferred to a petri dish inoculated with Staphylococcus aureus ATCC25923 in an amount of 108～109CFU/mL to simulate a Staphylococcus aureus infection. The case of the intervention (probiotic and/or HMO) added in the infection phase for each group was the same as for the culture phase, respectively. Two controls were used, respectively a condition without the pathogenic bacteria (nematode dishes containing E.coli OP50) and a condition with the pathogenic bacteria Staphylococcus aureus infection without any intervention (dishes containing only Staphylococcus aureus). The group of the culture stage added with the intervention substance without any intervention substance in the infection stage and the group of the culture stage added with no intervention substance in the infection stage are also set.
After several days of incubation, the nematodes were counted daily for their survival. If the nematode does not respond to the platinum filament, it can be considered dead. Two independent determinations were performed for each condition.
Statistical comparative analysis was performed on survival curves, and log rank survivval significance analysis was performed using the GraphPad Prism 4 statistical software package. Significant differences in survival on the third day of infection were analyzed by One-way ANOVA. Survival data for each day was statistically analyzed using Two-way ANOVA and significance was analyzed using Dunnett's posthoc test for each group.
Results of the experiment
As shown in FIG. 1, 15mg/mL of HMO 2' -FL and YLGB-1496 (10)8CFU) has obvious protection and promotion effects on the survival rate of the nematodes infected by staphylococcus aureus (P)<0.0001), and the protective effects of the two are relatively similar. And 107YLGB-1496 strain of (A)The survival rate of the nematodes also has better protection effect, although the effect is obviously lower than 108Survival Rate under circumstances (P)<0.05). When 2' -FL and YLGB-1496 strain (10)8CFU) to form a composition, in which case the survival rate of nematodes infected with staphylococcus aureus is significantly better than either of the two substances alone, i.e. either the strain alone or the HMO alone, and the 22% survival rate improvement (P) is achieved with the composition compared to the starting material alone<0.001). It can be seen that when 2' -FL and YLGB-GB1496 form a composition, the two show better synergistic effect.

Claims (9)

1. Application of composition of bifidobacterium longum subsp. infantis and breast milk oligosaccharide in preparation of formula milk powder capable of effectively improving resistance of organism to staphylococcus aureus infection and improving innate immunity and/or anti-aging effect of organism, wherein each 100g of formula milk powder contains 1 x 10 of bifidobacterium longum subsp. infantis3CFU～1×1012CFU, 0.1g to 10g of breast milk oligosaccharide.
2. The use according to claim 1, wherein the Bifidobacterium longum subsp.
3. The use of claim 1, wherein the total protein content of the formula milk powder is 10-20 g/100g, and the proportion of whey protein in the total protein is 40-70%; the fat content is 15 g-30 g/100 g; the carbohydrate content is 50 g-70 g/100 g.
4. The use of claim 1 or 2, wherein the formula milk powder contains 2700-4500 mg/100g linoleic acid and 270-450 mg/100g alpha-linolenic acid.
5. Use according to claim 1, wherein the formula is an infant formula or a non-infant formula.
6. The use of claim 1, wherein the formula further comprises one or more of nutrients, dietary fibers, inositol, taurine, L-carnitine, docosahexaenoic acid, arachidonic acid, lutein, nucleotides, lactoferrin.
7. Use according to claim 1, wherein the breast milk oligosaccharides comprise one or more of 2 '-fucosyllactose, 3' -fucosyllactose, lacto-N-tetraose, 3 '-sialyllactose, 6' -sialyllactose.
8. The use of claim 1, wherein the enhancing the resistance of an organism to infection by staphylococcus aureus comprises: improving the ability of an individual to prevent staphylococcus aureus infection, reducing the ability of an individual to be infected with staphylococcus aureus, and/or relieving food poisoning, enteritis, pneumonia, skin infection, wound ulceration and/or meningitis of the individual caused by staphylococcus aureus infection.
9. A formula milk powder with the effects of effectively improving the resistance of organisms to staphylococcus aureus infection and improving the innate immunity and/or anti-aging effect of the organisms is prepared by adding bifidobacterium longum subsp. infantis and breast milk oligosaccharide into the formula milk powder according to the application of any one of claims 1 to 8.
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